C-Terminal Extension of a Plant Cryptochrome Dissociates from the β-Sheet of the Flavin-Binding Domain

2021 ◽  
pp. 5558-5563
Author(s):  
Lukas Goett-Zink ◽  
Anna Lena Toschke ◽  
Jan Petersen ◽  
Maria Mittag ◽  
Tilman Kottke
Keyword(s):  
Binding Domain ◽  
Flavin Binding ◽  
Β Sheet

scholarly journals Blakeslea trispora Photoreceptors: Identification and Functional Analysis

2020 ◽  
Vol 86 (8) ◽  
Author(s):  
Wei Luo ◽  
Chao Xue ◽  
Yuzheng Zhao ◽  
Huili Zhang ◽  
Zhiming Rao ◽  
...  
Keyword(s):  
Large Scale ◽  
Binding Domain ◽  
Site Directed Mutagenesis ◽  
Flavin Mononucleotide ◽  
Blakeslea Trispora ◽  
Scale Production ◽  
Bioinformatics Analyses ◽  
Content Type ◽  
Flavin Binding

ABSTRACT Blakeslea trispora is an industrial fungal species used for large-scale production of carotenoids. However, B. trispora light-regulated physiological processes, such as carotenoid biosynthesis and phototropism, are not fully understood. In this study, we isolated and characterized three photoreceptor genes, btwc-1a, btwc-1b, and btwc-1c, in B. trispora. Bioinformatics analyses of these genes and their protein sequences revealed that the functional domains (PAS/LOV [Per-ARNT-Sim/light-oxygen-voltage] domain and zinc finger structure) of the proteins have significant homology to those of other fungal blue-light regulator proteins expressed by Mucor circinelloides and Neurospora crassa. The photoreceptor proteins were synthesized by heterologous expression in Escherichia coli. The chromogenic groups consisting of flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN) were detected to accompany BTWC-1 proteins by using high-performance liquid chromatography (HPLC) and fluorescence spectrometry, demonstrating that the proteins may be photosensitive. The absorbance changes of the purified BTWC-1 proteins seen under dark and light conditions indicated that they were light responsive and underwent a characteristic photocycle by light induction. Site-directed mutagenesis of the cysteine residual (Cys) in BTWC-1 did not affect the normal expression of the protein in E. coli but did lead to the loss of photocycle response, indicating that Cys represents a flavin-binding domain for photon detection. We then analyzed the functions of BTWC-1 proteins by complementing btwc-1a, btwc-1b, and btwc-1c into the counterpart knockout strains of M. circinelloides for each mcwc-1 gene. Transformation of the btwc-1a complement into mcwc-1a knockout strains restored the positive phototropism, while the addition of btwc-1c complement remedied the deficiency of carotene biosynthesis in the mcwc-1c knockout strains under conditions of illumination. These results indicate that btwc-1a and btwc-1c are involved in phototropism and light-inducible carotenogenesis. Thus, btwc-1 genes share a conserved flavin-binding domain and act as photoreceptors for control of different light transduction pathways in B. trispora. IMPORTANCE Studies have confirmed that light-regulated carotenogenesis is prevalent in filamentous fungi, especially in mucorales. However, few investigations have been done to understand photoinduced synthesis of carotenoids and related mechanisms in B. trispora, a well-known industrial microbial strains. In the present study, three photoreceptor genes in B. trispora were cloned, expressed, and characterized by bioinformatics and photoreception analyses, and then in vivo functional analyses of these genes were constructed in M. circinelloides. The results of this study will lead to a better understanding of photoreception and light-regulated carotenoid synthesis and other physiological responses in B. trispora.

Crystal Structure ofParacoccus denitrificansElectron Transfer Flavoprotein:  Structural and Electrostatic Analysis of a Conserved Flavin Binding Domain†,‡

Biochemistry ◽  
1999 ◽  
Vol 38 (7) ◽  
pp. 1977-1989 ◽  
Author(s):  
David L. Roberts ◽  
Denise Salazar ◽  
John P. Fulmer ◽  
Frank E. Frerman ◽  
Jung-Ja P. Kim
Keyword(s):  
Crystal Structure ◽  
Binding Domain ◽  
Flavin Binding

scholarly journals The basic keratin 10-binding domain of the virulence-associated pneumococcal serine-rich protein PsrP adopts a novel MSCRAMM fold

Open Biology ◽  
2014 ◽  
Vol 4 (1) ◽  
pp. 130090 ◽  
Author(s):  
Tim Schulte ◽  
Jonas Löfling ◽  
Cecilia Mikaelsson ◽  
Alexey Kikhney ◽  
Karina Hentrich ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Invasive Disease ◽  
Binding Domain ◽  
Upper Respiratory Tract ◽  
Binding Region ◽  
Sequence Identity ◽  
Novel Variant ◽  
Upper Respiratory ◽  
Β Sheet ◽  
Specific Virulence

Streptococcus pneumoniae is a major human pathogen, and a leading cause of disease and death worldwide. Pneumococcal invasive disease is triggered by initial asymptomatic colonization of the human upper respiratory tract. The pneumococcal serine-rich repeat protein (PsrP) is a lung-specific virulence factor whose functional binding region (BR) binds to keratin-10 (KRT10) and promotes pneumococcal biofilm formation through self-oligomerization. We present the crystal structure of the KRT10-binding domain of PsrP (BR 187–385 ) determined to 2.0 Å resolution. BR 187–385 adopts a novel variant of the DEv-IgG fold, typical for microbial surface components recognizing adhesive matrix molecules adhesins, despite very low sequence identity. An extended β-sheet on one side of the compressed, two-sided barrel presents a basic groove that possibly binds to the acidic helical rod domain of KRT10. Our study also demonstrates the importance of the other side of the barrel, formed by extensive well-ordered loops and stabilized by short β-strands, for interaction with KRT10.

Crystallographic Study of the Recombinant Flavin-Binding Domain of Baker's Yeast Flavocytochromeb2:  Comparison with the Intact Wild-Type Enzyme†

Biochemistry ◽  
2002 ◽  
Vol 41 (13) ◽  
pp. 4264-4272 ◽  
Author(s):  
L. M. Cunane ◽  
J. D. Barton ◽  
Z.-W. Chen ◽  
F. E. Welsh ◽  
S. K. Chapman ◽  
...  
Keyword(s):  
Baker’S Yeast ◽  
Binding Domain ◽  
Baker's Yeast ◽  
Wild Type ◽  
Wild Type Enzyme ◽  
Crystallographic Study ◽  
Flavin Binding

scholarly journals Probing nucleotide-binding effects on backbone dynamics and folding of the nucleotide-binding domain of the sarcoplasmic/endoplasmic-reticulum Ca2+-ATPase

2004 ◽  
Vol 379 (2) ◽  
pp. 235-242 ◽  
Author(s):  
Mona ABU-ABED ◽  
Oscar MILLET ◽  
David H. MacLENNAN ◽  
Mitsuhiko IKURA
Keyword(s):  
Endoplasmic Reticulum ◽  
Fluorescence Spectroscopy ◽  
Tryptophan Fluorescence ◽  
Nucleotide Binding ◽  
Binding Domain ◽  
Backbone Dynamics ◽  
Global Fold ◽  
Enhanced Mobility ◽  
Relaxation Experiments ◽  
Β Sheet

In muscle cells, SERCA (sarcoplasmic/endoplasmic-reticulum Ca2+-ATPase) plays a key role in restoring cytoplasmic Ca2+ levels to resting concentrations after transient surges caused by excitation–coupling cycles. The mechanism by which Ca2+ is translocated to the lumen of the ER (endoplasmic reticulum) involves major conformational rearrangements among the three cytoplasmic domains: actuator (A), nucleotide-binding (N) and phosphorylation (P) domains; and within the transmembrane Ca2+-binding domain of SERCA. CD, fluorescence spectroscopy and NMR spectroscopy were used in the present study to probe the conformation and stability of the isolated N domain of SERCA (SERCA-N), in the presence and absence of AMP-PNP (adenosine 5´-[β,γ-imido]triphosphate). CD and tryptophan fluorescence spectroscopy results established that the effects of nucleotide binding were not readily manifested on the global fold and structural stability of SERCA-N. 15N-backbone-relaxation experiments revealed site-specific changes in backbone dynamics that converge on the central β-sheet domain. Nucleotide binding produced diverse effects on dynamics, with enhanced mobility observed for Ile369, Cys420, Arg467, Asp568, Phe593 and Gly598, whereas rigidifying effects were found for Ser383, Leu419, Thr484 and Thr532. These results demonstrate that the overall fold and backbone motional properties of SERCA-N remained essentially the same in the presence of AMP-PNP, yet revealing evidence for internal counter-balancing effects on backbone dynamics upon binding the nucleotide, which propagate through the central β-sheet.

scholarly journals A novel mode of DNA recognition by a β-sheet revealed by the solution structure of the GCC-box binding domain in complex with DNA

1998 ◽  
Vol 17 (18) ◽  
pp. 5484-5496 ◽  
Author(s):  
Mark D. Allen ◽  
Kazuhiko Yamasaki ◽  
Masaru Ohme-Takagi ◽  
Masaru Tateno ◽  
Masashi Suzuki
Keyword(s):  
Solution Structure ◽  
Dna Recognition ◽  
Binding Domain ◽  
Gcc Box ◽  
Β Sheet

scholarly journals Fumarate Reductase and Succinate Oxidase Activity ofEscherichia coliComplex II Homologs Are Perturbed Differently by Mutation of the Flavin Binding Domain

2006 ◽  
Vol 281 (16) ◽  
pp. 11357-11365 ◽  
Author(s):  
Elena Maklashina ◽  
Tina M. Iverson ◽  
Yelizaveta Sher ◽  
Violetta Kotlyar ◽  
Juni Andréll ◽  
...  
Keyword(s):  
Oxidase Activity ◽  
Binding Domain ◽  
Fumarate Reductase ◽  
Flavin Binding ◽  
Succinate Oxidase

scholarly journals The Photocycle of a Flavin-binding Domain of the Blue Light Photoreceptor Phototropin

2001 ◽  
Vol 276 (39) ◽  
pp. 36493-36500 ◽  
Author(s):  
Trevor E. Swartz ◽  
Stephanie B. Corchnoy ◽  
John M. Christie ◽  
James W. Lewis ◽  
Istvan Szundi ◽  
...  
Keyword(s):  
Blue Light ◽  
Binding Domain ◽  
Blue Light Photoreceptor ◽  
Flavin Binding

scholarly journals Isolation and characterization of the flavin-binding domain of flavocytochrome b2 expressed independently in Escherichia coli

1995 ◽  
Vol 309 (2) ◽  
pp. 601-605 ◽  
Author(s):  
A Balme ◽  
C E Brunt ◽  
R L Pallister ◽  
S K Chapman ◽  
G A Reid
Keyword(s):  
Escherichia Coli ◽  
Binding Domain ◽  
Flavin Mononucleotide ◽  
Exponential Process ◽  
Flavocytochrome B2 ◽  
Terminal Domain ◽  
Isolation And Characterization ◽  
Flavin Binding ◽  
Single Exponential

Flavocytochrome b2 consists of two distinct domains. The N-terminal domain contains protohaem IX and the larger, C-terminal domain contains flavin mononucleotide (FMN). We describe here the isolation of the flavin-binding domain expressed in Escherichia coli independent of the cytochrome domain. The isolated domain is an efficient lactate dehydrogenase with ferricyanide as electron acceptor but reduces cytochrome c, the physiological oxidant for flavocytochrome b2, extremely poorly; electron transfer from the flavin-binding domain to the separately expressed cytochrome domain is undetectable. FMN reduction by lactate occurs as a single exponential process in the isolated flavin-binding domain, in contrast to the biphasic kinetics observed with native flavocytochrome b2.

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